全文获取类型
收费全文 | 71874篇 |
免费 | 6007篇 |
国内免费 | 5521篇 |
出版年
2023年 | 1443篇 |
2022年 | 1837篇 |
2021年 | 2701篇 |
2020年 | 2664篇 |
2019年 | 3933篇 |
2018年 | 3067篇 |
2017年 | 2349篇 |
2016年 | 2167篇 |
2015年 | 2360篇 |
2014年 | 4133篇 |
2013年 | 5109篇 |
2012年 | 2828篇 |
2011年 | 3478篇 |
2010年 | 2501篇 |
2009年 | 3114篇 |
2008年 | 3197篇 |
2007年 | 3489篇 |
2006年 | 3103篇 |
2005年 | 2555篇 |
2004年 | 2169篇 |
2003年 | 2040篇 |
2002年 | 1801篇 |
2001年 | 1464篇 |
2000年 | 1237篇 |
1999年 | 1102篇 |
1998年 | 1047篇 |
1997年 | 927篇 |
1996年 | 905篇 |
1995年 | 931篇 |
1994年 | 882篇 |
1993年 | 776篇 |
1992年 | 787篇 |
1991年 | 787篇 |
1990年 | 557篇 |
1989年 | 584篇 |
1988年 | 532篇 |
1987年 | 529篇 |
1986年 | 475篇 |
1985年 | 760篇 |
1984年 | 983篇 |
1983年 | 772篇 |
1982年 | 856篇 |
1981年 | 681篇 |
1980年 | 661篇 |
1979年 | 592篇 |
1978年 | 470篇 |
1977年 | 417篇 |
1976年 | 388篇 |
1974年 | 252篇 |
1973年 | 272篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
331.
A method is proposed for predicting the adjacency order in which strands pack in a -sheet in a protein, on the basis of its amino acid sequence alone. The method is based on the construction of a predicted contact map for the protein, in which the probability that various residue pairs are close to each other is computed from statistically determined average distances of residue pairs in globular proteins of known structure. Compact regions, i.e., portions of the sequence with many interresidue contacts, are determined on the map by using an objective search procedure. The proximity of strands in a -sheet is predicted from the density of contacts in compact regions associated with each pair of strands. The most probable -sheet structures are those with the highest density of contacts. The method has been tested by computing the probable strand arrangements in a five-strand -sheet in five proteins or protein domains, containing 62–138 residues. Of the theoretically possible 60 strand arrangements, the method selects two to eight arrangements as most probable; i.e., it leads to a large reduction in the number of possibilities. The native strand arrangement is among those predicted for three of the five proteins. For the other two, it would be included in the prediction by a slight relaxation of the cutoff criteria used to analyze the density of contacts. 相似文献
332.
Summary Lymph nodes contain an extensive array of extracellular matrix fibers frequently referred to as reticular fibers because of their reticular pattern and positive reaction with silver stains. These fibers are known to contain primarily type-III collagen. In the present study, frozen and plastic-embedded sections of mouse and human lymph nodes were subjected to immunostaining with a panel of monospecific antibodies directed against type-IV collagen, type-III collagen, laminin, entactin, and heparan sulfate proteoglycan. Immunofluorescent staining revealed that, in addition to being uniformly stained with antibodies to type-III collagen, these fibers also stained positively with antibodies to type-IV collagen and to other basement-membrane-specific components. Furthermore, the basement-membrane-specific antibodies stained the outer surface of individual fibers. These same type-III collagen-rich fibers were distinct from blood vascular basement membranes since they did not react with antibodies to factor VIII-related antigen, an endothelial-cell-specific marker. The role of these basement-membrane-specific components associated with the reticular fibers of lymphoid tissue is unknown. However, it is possible that the ligands promote attachment of reticular fibroblasts as well as macrophages and lymphocytes to the extracellular matrix fibers. 相似文献
333.
C. J. Duncan 《Cell and tissue research》1988,253(2):457-462
Summary This study compares the action of inhibitors of the eicosanoid cascade on calcium-induced myofilament damage in cardiac muscle of the perfused frog heart and incubated frog ventricle slices, and in skeletal muscle of incubated mammalian diaphragm and isolated and saponin-skinned amphibian pectoris cutaneous muscle. Mepacrine (10-5M) and indomethacin (3×10-6M) protected completely against myofilament damage induced by entry of calcium in the calcium-paradox in frog heart. However, inhibition of phospholipase A2 (PLA2) (with chlorpromazine, 2×10-4M, or mepacrine, 10-5M, 5x10-5M), of cyclo-oxygenase enzymes (with indomethacin, 3x10-6M to 10-5M or BW755C, 3.8x10-4M), or of lipoxygenase enzymes (with BW755C, 3.8x10-4M or nordihydroguaiaretic acid, 2x10-6M or 5x10-6M) all failed in intact cardiac or skeletal muscle cells to prevent the myofilament damage that is rapidly triggered by 10-2M caffeine, 6x10-6M ruthenium red, 10-4M DNP or 5 g ml-1 A23187. These agents also failed completely to protect against myofilament damage in saponin-skinned amphibian skeletal muscle when [Ca]i was raised to 8x10-6M. Thus, inhibition of PLA2 does not protect the myofilament apparatus against calcium released intracellularly, and it is suggested that mepacrine and indomethacin can block entry of calcium in the calcium-paradox in the amphibian heart. Chlorpromazine (2x10-4M) and mepacrine (10-3M) at zero [Ca] caused severe myofilament damage in skinned muscle, possibly due to an effect on membranes. Since inhibitors of PLA2 and of lipoxygenases prevent efflux of creatine kinase and sarcolemma damage in mammalian skeletal muscle, it is evident that experimentally-induced rises in [Ca]i (by caffeine or A23187) can trigger two separate pathways: (i) PLA2 and the arachidonic acid cascade which culminate in membrane damage, and (ii) a different, Ca-activated system that causes rapid damage of myofilaments. 相似文献
334.
Summary Ultrastructural studies of pancreatic islets have suggested that crinophagy provides a possible mechanism for intracellular degradation of insulin in the insulin-producing B-cells. In the present study, a quantitative estimation of crinophagy in mouse pancreatic islets was attempted by morphometric analysis of lysosomes containing immunoreactive insulin. Isolated islets were incubated in tissue culture for one week in 3.3, 5.5 or 28 mmol/l glucose. The lysosomes of the pancreatic B-cells were identified by morphological and enzyme-cytochemical criteria and divided into three subpopulations comprising primary lysosomes and insulin-positive or insulin-negative secondary lysosomes. Both the volume and numerical density of the primary lysosomes increased with increasing glucose concentration. The proportion of insulin-containing secondary lysosomes was highest at 5.5 and lowest at 3.3 mmol/l glucose. Insulin-negative secondary lysosomes predominated at 3.3 mmol/l glucose. Studies of the dose-response relationships of glucose-stimulated insulin biosynthesis and insulin secretion of the pancreatic islets showed that biosynthesis had an apparent Km-value for glucose of 7.0 mmol/l, whereas it was 14.5 mmol/l for secretion. The pronounced crinophagic activity at 5.5 mmol/l glucose may thus be explained by the difference in glucose sensitivity between insulin biosynthesis and secretion resulting in an intracellular accumulation of insulin-containing secretory granules. The predominance of insulin-negative secondary lysosomes at 3.3 mmol/l glucose may reflect an increased autophagy, whereas the predominance of primary lysosomes at 28 mmol/l glucose may reflect a generally low activity of intracellular degradative processes. 相似文献
335.
Dr. Yohichi Satoh 《Cell and tissue research》1988,253(2):397-402
Summary Previous studies have shown that the secretory products of Paneth cells contain antibacterial agents (lysozyme, IgA) that are affected by the bacterial milieu in the intestine. To investigate whether Paneth-cell secretion is controlled via cholinergic mechanisms, the ultrastructure of Paneth cells was studied in four animal groups: (1) germfree (GF) control mice (Jcl: ICR [GN], male, 13 weeks old), (2) GF mice injected subcutaneously with atropine sulfate (200 mg/kg body weight, dissolved in physiological saline 20 mg/ml), (3) ex-GF mice inoculated with feces from specific-pathogen-free (SPF) mice, and (4) ex-GF mice injected with atropine and inoculated with feces from SPF mice. In ex-GF mice inoculated with feces, 70–90% of the Paneth cells showed fewer secretory granules than those from GF mice (p<0.01). Approximately 30% of the Paneth cells had a large vacuole (3–10 m diameter) in the apical cytoplasm. Exocytosed electron-dense material from secretory granules was observed in a few crypt lumens. In ex-GF mice inoculated with feces and given atropine, about 90% of the Paneth cells contained numerous secretory granules, like those in GF control mice, but vacuolated Paneth cells and exocytotic figures were rare; thus the secretion of Paneth cells was blocked by atropine. It is therefore possible that the bacterial milieu in the intestine affects the secretory activity of Paneth cells via cholinergic mechanisms. 相似文献
336.
The lipopolysaccharides (LPS) of a rough (R) and a smooth (S) strain of Pseudomonas syringae pv. phaseolicola were analysed. The S-LPS revealed markedly more rhamnose and fucose, but less glucose, than the R-LPS. The presence of 3-O-methyl-rhamnose (acofriose) in the S-LPS was confirmed by cochromatography with authentic acofriose. SDS polyacrylamide gel electrophoresis of the S-LPS demonstrated a cluster of regularly spaced high molecular weight fractions, which was almost lacking in the R-LPS. The main fatty acids of the lipid A of both LPS species were 3-OH-10:0,3-OH-12:0,2-OH-12:0, and 12:0. Two N-linked diesters were demonstrated: 3-O(12:0)-12:0 and 3-O(2-OH-12:0)-12:0. S-LPS was subjected to mild hydrolysis and the degraded polysaccharide separated into three fractions by gel permeation chromatography on a Fractogel TSK HW-50 column. Fraction I, representing nearly only the O-specific side chain, consisted of rhamnose and fucose in a molar ratio of 4:1, with 4% of the rhamnose being 3-O-methylated (acofriose). Fraction II, representing mostly core material, was composed of glucose, rhamnose, heptose, glucosamine, galactosamine, alanine, and a still unidentified amino compound, in an approximate molar ratio of 3:1:1:1:1:1:1, and KDO. Fraction III consisted of released monomers and salts. The LPS was highly phosphorylated (3.28% phosphorus in the core fraction). The thus characterized composition of the LPS O-chain seems to be unique for the pathovar phaseolicola of P. syringae, although many similarities exist to other pathovars as well as to other bacterial species.Abbreviations LPS
lipopolysacchairdes
- GC/MS
combined gas liquid chromatography-mass spectrometry
- HVE
high voltage electrophoresis
- KDO
2-keto-3-deoxyoctonic acid
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecylsulfate
P.s. pv. phaseolicola is termed P. phaseolicola in the text 相似文献
337.
Summary Ventral thoracic neurosecretory cells (VTNCs) of the blowflies, Calliphora erythrocephala and C. vomitoria, innervating thoracic neuropil and the dorsal neural sheath of the thoracico-abdominal ganglion have been shown to be immunoreactive to a variety of mammalian peptide antisera. In the neural sheath the VTNC terminals form an extensive neurohaemal network that is especially dense over the abdominal ganglia. The same areas are invaded by separate, ut overlapping serotonin-immunoreactive (5-HT-IR) projections derived from neuronal cell bodies in the suboesophageal ganglion. Immunocytochemical studies with different antisera, applied to adjacent sections at the lightmicroscopic level, combined with extensive cross-absorption tests, suggest that the perikarya of the VTNCs contain co-localized peptides related to gastrin/cholecystokinin (CCK), bovine pancreatic polypeptide (PP), Met- and Leuenkephalin and Met-enk-Arg6-Phe7 (Met-enk-RF). Electron-microscopic immunogold-labeling shows that some of the terminals in the dorsal sheath react with several of the individual peptide antisera, whilst others with similar cytology are non-immunoreactive. In the same region, separate terminals with different cytological characteristics contain 5-HT-IR. Both 5-HT-IR and peptidergic terminals are localized outside the cellular perineurium beneath the acellular permeable sheath adjacent to the haemocoel. Hence, we propose that various bioactive substances may be released from thoracic neurosecretory neurons into the circulating haemolymph to act on peripheral targets. The same neurons may also interact by synaptic or modulatory action in the CNS in different neuropil regions of the thoracic ganglion. 相似文献
338.
To assess the dynamics of synthesis of the wall by regenerating Candida albicans protoplasts deposition of chitin and mannoproteins were investigated ultrastructurally using wheat germ agglutinin conjugated with either horseradish peroxidase or colloidal gold, and Concanavalin A coupled to ferritin respectively.Freshly prepared protoplasts lacked wheat germ agglutinin receptor sites but after 1–2 h of regeneration, they were detected. After 4–5 h of regeneration, the cell wall showed a discrete structure which was only labelled with wheat germ agglutinin in thin sections. At this stage of regeneration the outermost layer of the wall was labelled with clusters of Concanavalin A-ferritin particles.After 8 h regeneration, the cell wall appeared compact, and homogenously marked with wheat germ agglutinin whereas only the surface layers appeared consistently labelled with Concanavalin A-ferritin.From these observations we conclude that C. albicans protoplasts are able to regenerate in liquid medium a cell wall consisting of a network of chitin fibrils and mannoproteins at least (glucan polymers were not determined in the present cytological study). The former are the fundamental component of the inner layers at early stages of regeneration, whereas the latter molecules are predominant in the outer layers of the wall.Abbreviations WGA-HRP
wheat germ agglutinin conjugated with horseradish peroxidase
- WGA-Au
wheat germ agglutinin conjugated with colloidal gold
- Con A-ferritin
Concanavalin A coupled to ferritin 相似文献
339.
Morten Glasø Olav Hilmar Iversen Torstein Hovig 《Virchows Archiv. B, Cell pathology including molecular pathology》1988,56(1):221-235
The nature and significance of so-called dark keratinocytes in the epidermis during chemical carcinogenesis is still a matter
of concern and debate. Based on ultrastructural observations it has been suggested that dark cells most often are shrunken
cells. Reports on skin carcinogenesis, however, claim that dark cells are a sign of ongoing tumor promotion and represent
those stem cells in the epidermis from which the tumors originate. It is therefore important to find out whether these cells
are simply injured and shrunken cells, or vital cells of great importance for carcinogenesis. Dark cells are assumed to be
rich in ribosomes. There is evidence, however, that the observed number of dark cells is highly dependent on tissue fixation.
In the present ultrastructural study, morphometric methods were used to compare the effects of two different fixation procedures
on the amount of cytoplasmic ribosomes in dark cells from both untreated and carcinogen-treated hairless mouse epidermis.
The results show that the ultrastructural features of both dark and clear cells vary considerably with different fixation
procedures. In acetone-treated controls typical dark cells are only observed when the fixative has a lower osmotic activity
than the plasma. With iso-osmolal fixation typical dark cells are not observed. After an abortive two-stage carcinogenesis
treatment, in which a single application of 9,10-dimethyl-l,2-benzanthracene (DMBA) in acetone was followed by a single application
of 12-O-tetradecanoyl-13-acetate (TPA) in acetone, signs of cell injury could be found after both fixation procedures. With
DMBA/TPA and hypo-osmolal fixation the number of dark cells seemed to increase, whereas only signs of cell injury with occurrence
of some heavily altered “clear cells” dominated the picture with iso-osmolal fixation. Morphometry showed that both the numerical
and the volumetric densities of cytoplasmic ribosomes in basal keratinocytes varied most significantly with the fixation procedure
used. The cytoplasmic volumes did not vary in a way that could explain these differences. One might therefore assume that
the number of ribosomes depends on the fixative. Large swelling artifacts occurred when a fixative with low osmotic activity
was used, leading to compression of neighboring cells. Hence, an increased ribosomal density reported previously in dark cells
is probably related to such cell volume artifacts and does not reflect an actually increased quantity of ribosomes. With both
fixation procedures, a single application of DMBA followed by one of TPA appeared to produce an increased number of ribosomes
in basal keratinocytes. When hypo-osmolal fixation was used, however, treatment with DMBA/TPA did not influence the cytoplasmic
volume or the numerical density of ribosomes, in dark cells. This might indicate that so-called dark keratinocytes following
DMBA/TPA treatment are functionally inactive cells that appear more vulnerable than active cells to compression during hypo-osmolal
fixation. 相似文献
340.
在大鼠尾部,重复压力刺激皮肤感受野,当间隔时间小于2min时,多觉型伤害性感受器的单位放电数随间隔时间的缩短而减少。压力与辐射热交叉刺激同一感受野,随后刺激的放电数也显著减少。皮下注射致痛剂引起持续性放电的背景上,分别向感受野施加按压、辐射热或电针刺激,随着放电增多后出现一个放电减少的过程。刺激支配尾部的交感神经则使减少的放电显著增多。结果表明,多觉型伤害性感受器受到刺激兴奋后有个感受性降低的过程。本文讨论了这一过程在按摩、针灸缓解痛机制中的可能作用。 相似文献